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Characterization of Membrane Metal Threads by Proteomics and Analysis of A G Model CULHER-3372; No. of Pages 8 ARTICLE IN PRESS Journal of Cultural Heritage xxx (2017) xxx–xxx Available online at ScienceDirect www.sciencedirect.com Original article Characterization of membrane metal threads by proteomics and analysis of a 14th c. thread from an Italian textile a,b b b,∗ Aleksandra K. Popowich , Timothy P. Cleland , Caroline Solazzo a Department of Chemistry, University of Alberta, 11227 Saskatchewan Drive, Edmonton, Alberta T6G 2G2, Canada b Smithsonian’s Museum Conservation Institute, 4210 Silver Hill Road, Suitland, MD 20746, USA a r t i c l e i n f o a b s t r a c t Article history: Beginning in the 13th century, membrane metal threads – made out of animal skins (leather, parchment, Received 22 January 2018 and vellum) or membranous material (e.g., stomach, intestine) coated with metal – were the most popular Accepted 9 March 2018 variety of decorative metal threads used in European textiles. This work provides the proteomics ground- Available online xxx work for the identification of the species and the type of membrane used in the manufacture of a 14th century membrane gilded thread. A protocol for small sample extraction and nanoLC-Orbitrap MS/MS Keywords: analysis was first tested on standards of pig peritoneum and cow intestine metal-coated with or without Metal threads the presence of an egg adhesive. The proteomes of each membrane were characterized and compared Membrane by qualitative and quantitative bioinformatics; in addition to the predominant collagen proteins in each Collagen membrane type, minor tissue-specific proteins (e.g., smooth muscle proteins from intestine standards) Cyprus gold Proteomics were detected. Species-specific collagen peptides (i.e., from collagen I and collagen III) were confidently nanoLC-Orbitrap MS/MS identified to determine the species of origin, regardless of the application of metal and egg-based adhe- sives. Likewise, the thin layer of egg adhesive was successfully characterized with the detection of egg white (ovalbumin, ovotransferrin, lysozyme) and egg yolk (vitellogenin I, II, III) proteins. When applied to the thread from a 14th century Italian textile, this comprehensive methodology resulted in the iden- tification of seven collagen I and III peptides specific to cow, as well as other proteins suggesting that the ancient thread was made with intestine or stomach membrane without the use of an egg-based adhesive. © 2018 Elsevier Masson SAS. All rights reserved. 1. Introduction or flattened strips used directly in weaving, III. Metal wire or strips wound around a fiber core, IV. Metallic surface applied to organic Decorative metal threads have been extensively used for the wrappings (cellulosic or proteinaceous) wound around a fiber core, embellishment of textiles since ancient times. Many examples of and V. Metallic surface applied to organic strips (cellulosic or pro- metal threads exist in artifacts of cultural importance, and even teinaceous) without a fiber core [2–6]. earlier references to lavish gold and silver textiles can be found in Protein metal threads (Categories IV and V) were made from ancient texts, including a description of an ephod containing gold membranous tissues (e.g., stomach or intestinal walls of animals), in the Old Testament of the Bible (Exodus 39:2–3 “They hammered although skin has also been used as a substrate [2,7]. Research at the out thin sheets of gold and cut strands (. .)”). The popularity of end of the 19th century suggested that gilt membranes were made textiles woven or embroidered with metal threads persisted, and using the intestines from slaughtered animals [8]. Cow intestine their use is frequently associated with textiles intended to portray was similarly used in the manufacture of gold foil, also called gold- wealth or symbolic importance [1]. Metal threads were most often beater’s skin, during the same time period. Metal was applied to the made of gold, silver, or their alloys, often gilt, with the fabrication membrane with metal leaves or by mottle gilding, using either the method differing by region and changing over time [2]. Five cate- natural exudates of the organic membrane or an additional adhe- gories describing the use of metals in textiles have been defined: I. sive [7]. Reports have described the use of egg white, egg yolk, Metal applied with adhesive to already woven fabrics, II. Metal wire animal fat, animal and fish glues, gums, and clays as adhesives but to date, no scientific investigation has been carried to substantiate the presence and nature of the adhesives [4,7,9]. In category IV threads, also known as gilt membranes or Cyprus gold, the gilded membrane ∗ was cut into thin strips and wound around a silk or linen core [2]. No Corresponding author. E-mail address: [email protected] (C. Solazzo). adhesive was used between the gilded organic wrapping and the https://doi.org/10.1016/j.culher.2018.03.007 1296-2074/© 2018 Elsevier Masson SAS. All rights reserved. Please cite this article in press as: A.K. Popowich, et al., Characterization of membrane metal threads by proteomics and analysis of a 14th c. thread from an Italian textile, Journal of Cultural Heritage (2017), https://doi.org/10.1016/j.culher.2018.03.007 G Model CULHER-3372; No. of Pages 8 ARTICLE IN PRESS 2 A.K. Popowich et al. / Journal of Cultural Heritage xxx (2017) xxx–xxx Table 1 fiber core; instead, the twist of the metal membrane thread around Reference standards. the core was sufficient to keep the material in place. Migration of adhesive and wrapping material into the core fiber has however Reference Membrane Adhesive Fixation Metal leaf type been observed [5]. In category V threads the metal was applied to Pig untreated Peritoneum None None None membrane strips and used in textiles without being wound [4]. Pig gilt Peritoneum None None Gilt silver Membrane metal threads were first used in the 11th century by Pig gilt heat Peritoneum None Heat Gilt silver Pig gilt egg white Peritoneum Egg white None Gilt silver Levantine traders in Cyprus [1]. Beginning in the 13th century, gilt Pig gilt egg yolk Peritoneum Egg yolk None Gilt silver membranes became the most common variety of metal threads Cow untreated Gut None None None used in European textiles, especially those from Italy and Spain, Cow silver Gut None None Silver as well as Western Asia because they were flexible, lightweight, Cow silver heat Gut None Heat Silver Cow silver egg white Gut Egg white None Silver and inexpensive [7]. Substantial focus has been given to identi- fying the metals used in metal threads, as studies show that the composition of the metal and gold/silver ratios are suggestive of geographic origin of the metal thread [6,7,10]. Recently, a range of electron microscopy techniques and micro-Raman spectroscopy have been used to characterize cross-sections of metal threads revealing details of the production technology through study of the 3D texture [11]. Conversely, little attention has been given to identifying the membrane type or animal species used in organic metal threads, although the elucidation of this information would provide valuable insight into the origin, production technique, and use of membrane metal threads in the medieval period [1]. Identi- fication of the membrane portion of the substrate poses difficulty, especially when a textile is in a deteriorated state. Using mor- phology on the microscopic scale, differences between leather, membranous material, parchment, and vellum have been made [1,2]; however, the appearance of the substrate can be drastically altered from its original state because of decomposition, embrit- tlement, wear, and treatment damage, making visual identification indefinite and oftentimes impossible. In contrast to morphological identification, DNA amplification and molecular biology techniques have been used to identify the animal species in organic metal threads found in textiles from the 11–15th centuries [9]. Unfor- tunately, this study could only narrow down the identification to a few candidate species. Here, we have adopted a bottom-up pro- Fig. 1. 14th century Italian textile (#21714-1) from the History Museum in Graz, teomics approach to analyze a 14th c. membrane thread: the entire Austria, ©Universalmuseum Joanneum, Museum für Geschichte, Kulturhistorische Sammlung/Institute of Conservation, University of Applied Arts Vienna/Elisabeth membrane sample is characterized through a single extraction and Delvai 2017. The arrow points to where the thread was sampled. In the bottom right, digestion of the whole extract, including the membrane proteins the image of the thread was acquired with HIROX KH-8700 3D digital microscope and other binding proteins if present. The complex mixture is (Hirox-USA, Inc., NJ), courtesy of Thomas Lam (Smithsonian’s Museum Conservation separated by liquid chromatography before analysis by an Orbit- Institute). Software: PowerPoint 2013. rap Velos mass spectrometer and the data obtained are searched against a public database. In the absence of historical informa- thus revealing tissue-specific proteins, while peptides specific to tion on the membrane’s animal origin, any species can be targeted the membrane species and the egg adhesive were characterized through a proteomics database search. in untreated and treated samples. Finally, the developed protocol was applied to a membrane metal thread from a 14th century Ital- ian textile (Fig. 1), thought to be made from an animal’s internal 2. Research aims organ and not skin. To resolve the identification issue of the organic substrate of 3. Materials and methods a metal thread several centuries old, a proteomics approach was devised. Proteomics has been used successfully to identify binders 3.1. Standards [12] and collagen-based substrates such as parchment [13], but has never been used to characterize membrane metal threads. The The gilt membrane reference standards were prepared at the aim of this project is to show that a complex organic substrate, Universität für angewandte Kunst (University of Applied Arts composed of a membrane base and protein binders, can be com- Vienna, Austria) based on what is known of traditional protocols prehensively characterized, including the type and species of the [14,15].
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